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Recombinant scFv antibodies against infectious pancreatic necrosis virus isolated by flow cytometry

文献类型： 外文期刊

作者： Xu, Li-Ming 1 ; Zhao, Jing-Zhuang 1 ; Liu, Miao 1 ; Cao, Yong-Sheng 1 ; Yin, Jia-Sheng 1 ; Liu, Hong-Bai 1 ; Lu, Tongy 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Heilongjiang River Fishery Res Inst, Harbin 150070, Peoples R China

关键词： Antibody library; Antibody-antigen co-expression; Bacterial display; Fluorescence activated cell sorter; Infectious pancreatic necrosis virus; ChRtm213; Single chain variable fragment

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2016 年 237 卷

页码：

收录情况： SCI

摘要： Infectious pancreatic necrosis is a significant disease of farmed salmonids in China. In this study, a single chain variable fragment (scFv) antibody library derived from rainbow trout (Oncorhynchus mykiss) and viral protein VP2 of a Chinese infectious pancreatic necrosis virus (IPNV) isolate ChRtm213 were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry (FCM) to select IPNV specific antibodies. Six antibody clones with different mean fluorescence intensities (MFI) were obtained by picking colonies at random. The antibody clones were expressed and purified. The purified IPNV-specific scFv antibodies were used successfully in Western blotting, enzyme linked immunosorbent assay (ELISA) and an immunofluorescence antibody test (IFAT). This method provides a high throughput means to screen an antibody library by flow cytometry, and isolate a panel of antibody that can be used as potential reagents for the detection and study of IPNV that are prevalent in China. (C) 2016 Elsevier B.V. All rights reserved.

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